Phenylbismuth bis(2-pyridinethiol 1-oxide)

ABSTRACT

PHENYLBISMUTH BIS(2-PYRIDINETHIOL 1-OXIDE), A NEW ANTIBACTERIAL AND ANTIFUNGAL COMPOUND, AND COMPOSITIONS CONTAINING SAME.

United States Patent 3,753,990 PHENYLBISMUTH BIS(2-PYRIDINETHIOL'l-OXIDE) John Downing Curry, Oxford, Ohio, assignor to The Procter &Gamble Company, Cincinnati, Ohio No Drawing. Continuation-impart ofabandoned application Ser. No. 98,086, Dec. 14, 1970. This applicationJan. 17, 1972, Ser. No. 218,584

Int. Cl. (307d 31/50 U.S. Cl. 260270 R 1 Claim ABSTRACT OF THEDISCLOSURE Phenylbismuth bis(2-pyridinethiol l-oxide), a newantibacterial and antifungal compound, and compositions containing same.

CROSS-REFERENCE TO RELATED APPLICATION This a continuation-in-part of mycopending U.S. patent application, S.N. 98,086 filed Dec. 14, 1970 forPhenylbismuth Di(Pyridinethione-N-Oxide) and Compositions ContainingSame now abandoned.

BACKGROUND OF THE INVENTION Field of the invention The present inventionrelates to a new compound, phenylbismuth bis(2-pyridinethiol l-oxide),which can be formed by the reaction between sodium 2-pyridinethiol 1-oxide and diphenylbismuth acetate. This invention also relates to theuse of this compound, which is highly efiective against a broad spectrumof bacteria as well as yeast and fungal species, especially when used onthe skin, where the compound is fairly substantive, and to compositions,including detergent compositions, containing said compound.

Prior art SUMMARY OF THE INVENTION Phenylbismuth bis(2pyridinethioll-oxide) has the formula:

N l o Compositions containing phenylbismuth bis(2-pyridinethiol l-oxide)have excellent broad-spectrum antibacterial and antifungal action.

Phenylbismuth bis(2-pyridinethiol l-oxide), hereinafter referred to asPBDP, is a yellow powder having a melting point at about 215-218". It isgenerally insoluble in most common solvents, but is soluble to theextent of about 1% in dimethylsulfoxide, dimethylformamide ordimethylacet- 3,753,990 Patented Aug. 21, 1973 ice amide. The dry solidPBDP is stable and the PBDP appears stable in the presence of soapsolutions (pH 9-10) and hydrochloric acid (pH 1).

Phenylbismuth bis(2-pyridinethiol l-oxide) has been shown to exhibitantibacterial and antifungal effectiveness against, e.g., Gram-positiveorganisms such as: Brevibacterium ammoniagenes, Erysipelothrixinsidiosa, Lact0- bacillus leichmanni, Bacillus subtilus,, Clostridiumperfringens, Staphylococcus aureus, Staphylococcus epidermiais, Sarcinalutea, Streptococcus faecalis, Cor ynebacterium acnes, Axilladiphtheriod #16, Oral Streptococcus #126, Mycobacterium phlei,Mycobactei'ium smegmatis, and Mycobacterium balnei; Gram-negativeorganisms such as: Alkaligenes faecalis, Salmonella typhosa, Shigellaflexneri, Mima polymvrpha, Herellea vaginicola, Hemophilus gallinarum,Proteus mirabilis, Vibrio metschnikovii, Neisseria flavescens,Escherichia coli, Pseudomonas aeruginosa, Serratia marcescens,Klebsiella pneumoniae, and Enterobacter acrogenes; and fungi (includingyeasts and dematophytes (such as: Aspergillus niger, Mycrosporum gypseumTrichophyton rubrum, T richophyton interdigitale, Saccharomycescerevisiae, Candida albicans, and Pityrosporum' o vale.

Because of the extremely broad antibacterial and antifungaleifectiveness of PBDP it is desirably used as a component of surgicalscrub products, bar soaps, shampoos, oral products (e.g., toothpastes,mouthwashes, etc.), first aid sprays, foot powder, deodorants(especially underarm deodorants), first aid creams, toilet bowlcleaners, hard surface cleaners, detergents, paints, cosmetics,spermicides, burn dressings, animal feeds, wood preservatives,mildewcides, germicides, algacides, fungicides, medicines, packingpreservatives, etc. In extremely small amounts, e.g., 10 ppm, the 'PBD'Pwill inhibit the growth of a broad variety of organisms. When the PBDPis incorporated in larger amounts, the compositions can be used todestroy organisms, as required. For example, in medicine, both forhumans and veterinary medicine, the PBDP can be used to treat a varietyof diseases and/or organisms including acne, leprosy psoriasis, warts,intestinal parasites, dandruif, pseudomonas or coli bacteria, fungus(e.g., athletes foot), vaginal infections, etc. Compositions containingPBDP can be used to treat tissue either to prevent infections or to cureinfections.

PBDP is not inactivated by urine or hair oil, but its activity issomewhat diminished by human serum or human red blood cell lysate. Itsactivity is somewhat greater in acid to neutral media against S. aureusand E. coli. PBDP is substantive to tooth enamel and cellulose. Thus,PBDP can be used to prevent the growth of plaque in the mouth and can beused against gingivitis. Also, it can be used on bandages, diapers, andsanitary napkins to prevent infections, diaper rash, odor, etc. PBDP canalso be used as a slimicide and preservative for paper and wood.

Preparation of phenylbismuth bis(2-pyridinethiol l-oxide) Phenylbismuthbis(2-pyridinethiol l-oxide) can be prepared by reacting diphenylbismuthacetate with sodium Z-pyridinethiol l-oxide in a 1:1 molar ratio in asolvent such as dimethyl formamide. The compound, a yellow precipitate,can then be filtered and purified as exemplified hereinafter in ExampleI. Other reactions to form phenylbismuth bis(2-pyridinethiol l-oxide)include the reaction of sodium, potassium, ammonium, or lithium 2-pyridinethiol 1 oxide or any other salt containing a loosely held cationwith a phenylbismuth dihalide (e.g., a chloride, bromide, or iodide).

Compositions containing phenylbismuth bis(2-pyridinethiol l-oxide)Phenylbismuth bis(2-pyridinethiol l-oxide) can be used in aqueous and/or non-aqueous solvents to provide antibacterial action. Preferably, thecompositions will contain from about 0.2% to about 3% phenylbismuthbis(2- pyridinethiol l-oxide), most preferably from about 1% to about2%. Desirable compositions are those containing soap and non-soapsynthetic detergent compounds. Preferably, the synthetic detergentcompounds are cationic, amphoteric, including zwitterionic or nonionic.

The term soap as used herein is meant to designate alkali metal soapssuch as the sodium and potassium salts of the higher fatty acids ofnaturally occurring plant or animal esters, e.g., palm oil, coconut oil,babassu oil, soybean oil, castor oil, tallow, whale and fish oils,grease and lard and mixtures thereof. Sodium and potassium soaps can bemade by direct saponification of the fats and oils or by theneutralization of the fatty acids which are prepared in a separatemanufacturing process. Examples of suitable soaps are the sodium,potassium, ammonium and alkylolammonium salts of higher fatty acids (C-C Particularly useful are the sodium and potassium salts of themixtures of fatty acids derived from coconut oil and tallow, i.e.,sodium or potassium tallow and coconut soap.

Anionic synthetic detergents which can be used with the antibacterial,antifungal, and antiyeast compound of the present invention can bebroadly defined as the watersoluble salts, including the alkali metal,ammonium and substituted ammonium salts, of organic sulfuric reactionproducts having in their molecular structure an alkyl radical containingfrom about 8 to about 22 carbon atoms and a radical selected from thegroup consisting of sulfonic acid and sulfuric acid ester radicals.

Important examples of the synthetic detergents which can be used withthe compound of the present invention are the following: alkali metal(e.g., sodium and potassium), ammonium and substituted ammonium (e.g.,lower alkyl ammonium) salts of the following: alkyl sulfates, especiallythose obtained by sulfating the higher alcohols produced by reducing theglycerides of tallow or coconut oil; random paraflin sulfonates, inwhich the alkyl group contains from about 8 to about 22 carbon atoms,prepared by treating random paraffin hydrocarbons in sulfur dioxide andchlorine in the presence of light followed by treating with a base;branched or linear alkyl benzene sulfonates, in which the alkyl groupcontains from about 8 to about 18 carbon atoms, preferably from about toabout 14 carbon atoms, especially those of the types described in US.Pat. Nos. 2,220,099, and 2,477,383; sodium alkyl glyceryl ethersulfonates, especially those ethers of the higher alcohols derived fromtallow and coconut oil; coconut oil fatty acid monoglyceride sulfatesand sulfonates; sulfuric acid esters of the reaction product of one moleof a higher fatty alcohol (e.g., tallow or coconut alcohols) and fromabout 1 to about 6, preferably about 3 moles of ethylene oxide; alkylphenol ethylene oxide ether sulfates with about 4 units of ethyleneoxide per molecule and in which the alkyl radicals contain about 9carbon atoms; the reaction product of fatty acids esterified withisethionic acid and neutralized with sodium hydroxide where, forexample, the fatty acids are derived from coconut oil; fatty acid amidesof the methyl tauriue in which the fatty acids, for example, are derivedfrom coconut oil; sulfonated olefins of US. Pat. No. 3,332,880; andothers known in the art, a number being specifically set forth in US.Pat. Nos. 2,486,921, 2,486,922 and 2,396,278.

The nonionic synthetic detergentswhich can be used with theantibacterial compound of the present invention may be broadly definedas compounds produced by the condensation of alkylene oxide groups(hydrophilic in nature) with an organic hydrophobic compound which maybe aliphatic or alkyl-aromatic in nature. The length of the hydrophilicor polyoxyalkylene radical which is condensed with any particularhydrophobic group can be readily adjusted to yield a water-solublecompound having the desired degree of balance between hydrophilic andhydrophobic elements.

For example, a well-known class of nonionic synthetic detergents is madeavailable on the market under the trade name of Pluronic. Thesecompounds are formed by condensing ethylene oxide with a hydrophobicbase formed by the condensation of propylene oxide with propyleneglycol. The hydrophobic portion of the molecule which, of course,exhibits water insolubility has a molecular weight of from about 1,500to about 1,800. The addition of polyoxyethylene radicals to thishydrophobic portion tends to increase the water solubility of themolecule as a whole and the liquid character of the products is retainedup to the point where polyoxyethylene content is about 50% of the totalweight of the condensation product.

Other suitable nonionic synthetic detergents include:

(1) The polyethylene oxide condensates of alkyl phenols, e.g., thecondensation products of alkyl phenols having an alkyl group containingfrom about 6 to 12 carbon atoms in either a straight chain or branchedchain configuration, with ethylene oxide, the said ethylene oxide beingpresent in amounts equal to 10 to 60 moles of ethylene oxide per mole ofalkyl phenol. The alkyl substituent in such compounds may be derivedfrom polymerized propylene, diisobutylene, octane, or nonane, forexample.

(2) Those derived from the condensation of ethylene oxide with theproduct resulting from the reaction of propylene oxide and ethylenediamine-products which may be varied in composition depending upon thebalance between the hydrophobic and hydrophilic elements which isdesired. For example, compounds containing from about 40% to aboutpolyoxyethylene by weight and having a molecular weight of from about5,000 to about 11,000 resulting from the reaction of ethylene oxidegroups with a hydrophobic base constituted of the reaction product ofethylene diamine and excess propylene oxide, said base having amolecular weight of the order of 2,500 to 3,000, are satisfactory.

(3) The condensation product of aliphatic alcohols having from 8 to 18carbon atoms, in either straight chain or branched chain configuration,with ethylene oxide, e.g., a coconut alcohol ethylene oxide condensatehaving from 10 to 30 moles of ethylene oxide per mole of coconutalcohol, the coconut alcohol fraction having from 10 to 14 carbon atoms.

(4) Long chain tertiary amine oxides corresponding to the followinggeneral formula, R R R N 0, wherein R contains an alkyl, alkenyl ormonohydroxy alkyl radical of from about 8 to about 18 carbon atoms, from0 to about 10 ethylene oxide moieties, and from 0 to 1 glyceryl moiety,and R and R contain from 1 to about 3 carbon atoms and from 0 to about 1hydroxy group, e.g., methyl, ethyl, propyl, hydroxy ethyl, or hydroxypropyl radicals. The arrow in the formula is a conven tionalrepresentation of a semi-polar bond. Examples of amine oxides suitablefor use in this invention include dimethyldodecylamine oxide, oleyldi(2hydroxyethyl)- amine oxide, dimethyloctylamine oxide,dimethyldecylamine, oxide, dimethyltetradecylamine oxide,3,6,9-trioxaheptadecyldiethylamine oxide, di(2hydroxyethyl)tetradecylamine oxide, Z-dodecoxyethyldimethylamine oxide,3 dodecoxy-Z-hydroxypropyldi(3-hydroxypropyl)amine oxide,dimethylhexadecylamine oxide.

(5) Long chain tertiary phosphine oxides corresponding to the followinggeneral formula RRR"P- O, wherein R contains an alkyl, alkenyl ormonohydroxyalkyl radical ranging from 8 to 18 carbon atoms in chainlength, from 0 to about 10 ethylene oxide moieties and from 0 to lglyceryl moiety and R and R" are each alkyl or monohydroxyalkyl groupscontaining from 1 to 3 carbon atoms. The arrow in the formula is aconventional representation of a semi-polar bond. Examples of suitablephosphine oxides are:

dodecyldimethylphosphine oxide, tetradecyldimethylphosphine oxide,tetradecylmethylethylphosphine oxide,3,6,9-trioxaoctadecyldimethylphosphine oxide, cetyldimethylphosphineOxide, 3-dodecoxy-2-hydroxypropyldi (Z-hydroxyethyl phosphine oxide,stearyldimethylphosphine oxide, cetylethylpropylphosphine oxide,oleyldiethylphosphine oxide, dodecyldiethylphosphine oxide,tetradecyldiethylphosphine oxide, dodecyldipropylphosphine oxide,dodecyldi(hydroxymethyl)phosphine oxide,dodecyldi(2-hydroxyethyl)phosphine oxide,tetradecylmethyl-Z-hydroxypropylphosphine oxide, oleyldimethylphosphineoxide, 2-hydroxydodecyldimethylphosphine oxide.

(6) Long chain dialkyl sulfoxides containing one short chain alkyl orhydroxy alkyl radical of 1 to about 3 carbon atoms (usually methyl) andone long hydrophobic chain which contains alkyl, alkenyl, hydroxy alkyl,or keto alkyl radicals containing from about 8 to about 20 carbon atoms,from to about ethylene oxide moieties and from 0 to l glyceryl moiety.Examples include:

octadecyl methyl sulfoxide,

2-ketotridecyl methyl sulfoxide, 3,6,9-trioxaoctadecyl 2-hydroxyethylsulfoxide, dodecyl methyl sulfoxide,

oleyl 3-hydroxypropyl sulfoxide,

tetradecyl methyl sulfoxide, S-methoxytridecyl methyl sulfoxide3-hydroxytridecyl methyl sulfoxide, 3-hydroxy-4-dodecoxybutyl methylsulfoxide.

The zwitterionic synthetic detergents useful with the antibacterialagent of the present invention can be broadly described as derivativesof aliphatic quaternary ammonium, phosphonium, and sulfonium compounds,in which the aliphatic radicals can be straight chain or branched, andwherein one of the aliphatic substituents contains from about 8 to 18carbon atoms and one contains an anionic water solubilizing group, e.g.,carboxy, sulfonate, sulfate, phosphate, or phosphonate. A generalformula for these compounds is:

wherein R contains an alkyl, alkenyl, or hydroxy alkyl radical of fromabout 8 to about 18 carbon atoms, from 0 to about 10 ethylene oxidemoieties and from 0 to l glyceryl moiety; Y is selected from the groupconsisting of nitrogen, phosphorus, and sulfur atoms; R is an alkyl ormonohydroxyalkyl group containing 1 to about 3 carbon atoms; x is 1 whenY is a sulfur atom and 2 when Y is a nitrogen or phosphorus atom, R isan alkylene or hydroxyalkylene of from 1 to about 4 carbon atoms and Zis a radical selected from the group consisting of carboxylate,sulfonate, sulfate, phosphonate, and phosphate groups.

Examples include:

4- [N,N-di Z-hydroxyethyl -N-octadecylammonio] butane-l-carboxylate; 5-[S-3-hydroxypropyl-S-hexadecylsulfonio] 3-hydroxypentane-l-sulfate;3-[P,P-diethyl-P-3,6,9-trioxatetradecoxylphosphonio]-2-hydroxy-propane-l-phosphate; 3- [N,N-dipropyl-N-3-dodecoxy-2-hydroxypropylammonio]-propane-l-phosphonate;3-(N,N-dimethyl-N-hexadecylammonio)propane-1- sulfonate;

3- (N,N-dimethyl-Nhexadecylammonio -2-hydroxypropane-l-sulfonate;

4- [N,N-di (Z-hydroxyethyl) -N- (2-hydroxydodecyl) ammonio-butanel-carboxylate;

3- [S-ethyl-S- 3-dodecoxy-2-hydroxypropyl sulfonio] propanel-phosphate;

3- [P,P-dimethyl-P-dodecylphosphonio]-propane-1- phosphonate and 5-[N,N-di 3-hydroxypropyl -N-hexadecylammonio] -2- hydroxy-pentanel-sulfate.

The amphoteric synthetic detergents useful in the present invention canbe broadly described as derivatives of aliphatic secondary and tertiaryamines in which the aliphatic radical can be straight chain or branchedand wherein one of the aliphatic substitutents contains from about 8 toabout 18 carbon atoms and one contains an anionic water solubilizinggroup, e.g., carboxy, sulfonate, sulfate, phosphate, or phosphonate.Examples of compounds falling within this definition are sodium3-dodecylaminopropionate, sodium 3-dodecylaminopropane sulfonate,dodecyl-fi-alanine, N-alkyl-taurines such as the one prepared byreacting dodecylamine with sodium isethionate according to the teachingof US. Pat. No. 2,658,072, N-higher alkyl aspartic acids such as thoseproduced according to the teaching of US. Pat. No. 2,438,091, and theproducts sold under the trade name Miranol and described in U.S. Pat.No. 2,528,378.

Cationic synthetic detergents include those quaternary ammonium,quaternary phosphonium, and ternary sulfonium compounds containing asingle straight chain or branched aliphatic radical containing fromabout six to about 20 carbon atoms such as dodecyltrimethylammoniumchloride; nonylbenzylethyldimethylammonium nitrate; tetradecylpyridiniumbromide; octadecylbutylpropylmethylphosphonium nitrite;decyldimethylsulfonium chloride; etc.

Detergent formulations containing the antibacterial compositions of thepresent invention can also contain from about 0% to about 90%,preferably from about 10% to about 90% of water-soluble alkalinedetergency builder salts, either of the organic or inorganic types.Examples of such builder salts can be found in US. Pat. No. 3,336,233,issued Aug. 15, 1967, column 9, lines 2966, which is incorporated hereinby reference. However, at a very high pH, the compound may undergoalkaline hydrolysis and accordingly, for long-term stability, thedetergent formulations should have a pH of less than about 10 andpreferably the pH should be approximately neutral.

The detergent formulations can also contain any of the usual adjuvants,diluents, and additives, for example, perfumes, anti-tarnishing agents,anti-redeposition agents, dyes, fluorescers, suds builders, sudsdepressors and the like without detracting from the advantageousproperties of the antibacterial compositions of the present invention.

Examples of diluents which may be incorporated into a syntheticdetergent bar in amounts of up to about of the bar include soaps,especially alkaline earth metal insoluble soaps (alkaline earth salts ofhigher fatty acids); starches such as cornstarch; and clays such aschina clay or fullers earth. Other diluents include inorganic salts suchas sodium and potassium chlorides and sulfates. Such diluents add bulkto the bar and improve its cosmetic properties without impairing itsdetergent or cohesive properties.

The antibacterial and antifungal eflicacy of the phenylbismuthbis(2-pyridinethiol l-oxide) is disclosed in the following examples.

EXAMPLE 1 Preparation of phenylbismuth bis(2- pyridinethiol l-oxide)164.7 grams of diphenylbismuth acetate (.39 mole) were slurried in 2,600ml. of dimethylformamide. 59.7

grams of sodium Z-pyridinethiol l-oxide (.39' mole, 97.3%) weredissolved in 1,300 ml. of dimethylformamide and 18 g. of water wereadded to this solution. The two solutions were then mixed while stirringat about 32 C. The solution turned yellow. The solution was continuouslystirred for 15 minutes, and then put on a steam bath for 15 minuteswhere a temperature of 52 C. was attained. The solution was then put ona heating mantle for approximately 1 hour at about 52- 58 C. Thesolution was then filtered, and the filtrate separated withdichloromethane and water (a ratio of approximately 1:1:2). The toplayer which contained water and dimethylforrnamide was discarded. Thebottom layer was filtered and washed with water and ether. The resultingimpure phenylbismuth bis(2-pyridinethiol l-oxide) was dried in vacuo.

Approximately 350 g. of this phenylbisrnuth bis(2- pyridinethiolI-oxide) was purified as follows: 30 g. of the crude material wasdissolved in 2,000 ml. dimethylacetamide at about 60 C. The solution wasstirred for approximately 5 minutes, during which time the phenylbismuthbis(2-pyridinethiol l-oxide) was dissolved, leaving the impurities whichwere separated by filtering. The filtrate was then diluted with an equalvolume of water to precipitate out the phenylbismuth bis(2-pyridinethioll-oxide). The mixture was cooled in an ice bath to precipitate as muchmaterial as possible, and this. material was then separated byfiltering. The residue was washed with water and ether and dried in aVacuum to produce essentially pure phenylbismuth bis(2-pyridinethiol 1-oxide). This procedure was repeated until all 350 g. had been processed.The final weight of the material was approximately 332 g. of materialhaving a melting point of about 218 C. The percent of carbon, hydrogen,nitrogen, bismuth, and sulfur present in the phenylbismuthbis(2-pyridinethiol l-oxide) as calculated and as found by analysis aregiven hereafter.

The phenylbismuth bis(2-pyridinethio l-oxide) prepared according to theprocess of Example I was tested for antibacterial efiicacy in thefollowing tests:

(1) Minimal inhibitory concentration (MIC) in agar 10,000 ppm. stocksolutions of phenylbismuth bis(2- pyridinethiol l-oxide) were preparedin dirnethylformamide (DMF). The active was diluted to the desired testconcentration and added to 19-20 ml. of molten trypticase soy agar (TSA)to give a series of graded concentrations (p.p.m.). Thorough rotation ofplates assured adequate mixing of the sanitizer before agarsolidification. Appropriate control plates containing plain TSA wereineluded. All plates remained at room temperature overnight.

Trypicase soy broth cultures for each test organism (24 hour) werediluted in peptone water to give approxi mately 100 cells per 0.01 ml.The agar plates, marked in sectors, were inoculated by dropping one dropof each diluted culture from a Kline antigen microdropper onto theappropriate sector of agar. The drops were allowed to dry. The plateswere incubated 48 hours at 37 C. and then examined for growth. Theweakest concentration which completely inhibited growth of an organismwas the MIC. The results for the tests were as follows:

Gram-positive organisms:

Staphylococcus aureus (S. a.) Staphylococcus epidermidis (S. ep.)Streptococcus faecalis (St. 1'.) Streptococcus sp. (St. 126) Sarcinalutea (Sar. l.) Diphtheroid sp. (Dip. 16) Brevibacterium ammoniagenesErysipelothrix insidiosa 1 Lactobacillus leichmanni Bacillus subtilusClostridium perfringens Corynebacterium aches Axilla diphtheroid #16Oral Streptococcus #126 Mycobacterium phlei Mycobacterium smegmatisMycobacterium balnei Gram-negative organisms:

Pseudomonas aerugz'nosa (Ps. a.) Escherichia coil (E. c.) Serratiamarcescens (S. m.) Klebsiella pneumoniae (K. p.) Enterobacter aerogenes(E. a.) Alkaligenes faecalis Salmonella typhosa Shigella flexneri Mirnapolymorpha Herellea vaginicola Hemophilus gallz'narum Proteus mirabilisVibrio metschnikovii Nez'sserz'a flavescens NNm mmJ -b-h-h-hNNw IUlUtl-Nrml- A 7-day old culture of Corynebacterium acnes grown anaerobicallyon brain heart infusion agar plus 1% glu cose (BI-IIA+l% G) was washedfrom the slant with 10 ml. of peptone water and drop inoculated onplates containing varying concentrations of phenylbismuth bis(2-pyridinethiol l-oxide) in BHIA+1% G. The MIC for this organism was 5ppm. A similar test was run using Pityrosporum ovale with TSA plus 0.1%of a :5 ratio of oleiczpalmitic acid as the culture medium. The MIC was5 ppm.

Fungistatic tests were performed using a modified gradient platetechnique reported by Hunt and Sandham (Applied Microbiology 17#2:329-30, 1969). Results were as follows: Aspergillus niger5.8 p.p.m.;Microsporum gypseum-1 p.p.m.

(II) Human corneum disc diffusion test Cornified epidermis was obtainedfrom the plantar surfaces of human volunteers by cutting sheets ofuniform thickness (0.4 mm.) with a dermatome. Discs, 6 mm. in diameter,were cut from the sheets with a #2 cork borer. A 1% solution ofphenylbismuth bis(2-pyridineth-iol 1- oxide) was prepared by dissolvingthe material in DMF to make a 10,000 p.p.m. stock solution and addingthe required amount of this solution to a 2.5% solution of pHisoderm (acommercially available skin cleansing detergent). Twenty ml. of the 1%solution of phenylbismuth bis(2-pyridiuethiol l-oxide) in pHisoderm wasadded to a small screw-cap vial. A control vial was prepared containing20 ml. of the pHisoderm solution alone. Six skin discs were added toeach of the vials which then were mechanically agitated for 10 minutesin a 50 C. water bath. The vial contents were transferred to a smallbeaker where the liquid was removed by suction, and the discs wererinsed three times with 30 ml. of distilled water,

each rinse followed by suction removal of the rinse water. The discswere then dried for two hours on a paper towel covered with a plexiglassshield to prevent air contamination. The dry discs were placed on thesurface of 15 m1. of solidified TSA in a Petri dish (one test disc andone control/dish). A l:l0,000 dilution of each of the six test organismsin TSA at 50 C. was prepared. Each Petri dish was overlaid with 10 ml.of agar seeded with one of the organisms. After agar solidification, thedisheswere refrigerated overnight to allow diffusion of any sanitizeraway from the disc into the medium. The next day, the dishes were placedin a 37 C. incubator for 24 hours. The diameter of any zone which wasclear of bacterial growth was measured in mm. These measurementsincluded the 6 mm. diameter disc. A clear zone is indicative ofsanitizer retention on the skin after the washpHisoderm controlsolution.

(HI) Filter disc diffusion test A test which was similar to the skindisc diffusion test was run using filter discs and several types offabric discs to give an indication of how much sanitizer would remainattached to paper and cloth after rinsing. The results were as follows:

Diameter Clear zone of disc,

mm. S. a E. c. Pa. a

Filter paper 12. 7 34 25 19 Diaper (gauze) 7.0 29 15.5 8 Terrycloth 9.33 23 18 Untreated muslin- 7. 0 23. 15. o 9 Disheloth 8. 0 25. 5 17 12(IV) Handwashing test 1% phenylbismuth bis(2-pyridinethiol l-oxide) inpHisoderm was used by 12 subjects who washed their hands four times.These subjects were chosen at random from a large group of availablesubjects who use non-sanitizer products exclusively for personalhygiene. The regimented washings in the laboratory followed thisprocedure: The hands were moistened under 100 F. running tap water. Theproduct was squeezed from a plastic bottle into the palm; the amountapproximated the size of a ZS-cent piece. The product was thendistributed over the hands and the lather was worked for 90 seconds. Thehands were then rinsed under running tap water for 30 seconds. The fourexposures were spread over two days, and on the afternoon of the secondday, the subjects washed their hands four times using a blank bar soapcontaining no antibacterial agents. A fifth standardized washing wasthen done in a basin containing one liter of sterile distilled waterwith careful rinsing in this basin. Aliquots were taken from thethoroughly mixed basin contents, added to 30 ml. of sterile distilledwater in a membrane funnel, and passed through sterile membrane filterswith vacuum suction. The filters were then incubated on pads saturatedwith 2x concentrated trypticase soy broth plus horse serum and 1% Tween80 in plastic dishes for 48 hours at 37 C. Colonies appearing on thefilters were counted and these numbers multiplied by the appropriatedilution factor to determine the number of bacteria/ liter in the fifthbasin contents. The numbers per basin for each panelist were convertedto logarithms to minimize unusually high or low counts. These valueswere averaged to determine the mean log of the fifth basin bacterialcount. Calculation of the log percent reduction for each subject wasmade using the log of 1.3 10 or 6.1184, an average fifth basin bacterialcount obtained from more than 500 handwashings by non-antibacterial soapusers. These values were averaged to obtain the mean log percentreduction. The results were as follows:

Log percent reduction:

Mean log 4.7818

A second handwashing test in which the subjects used a measured amountof product (5 ml.) showed the following result:

Log percent reduction:

Mean log 97.48

These tests show that phenylbismuth bis(2-pyridinethiol l-oxide) in adetergent vehicle is effective against normal skin bacterial in vivounder conditions of actual use.

(V) Skin disc drop overlay test S. a J 1 E. c. 2 Ps. a 10 As can be seenfrom the above data, phenylbismuth bis (Z-pyridinethiol l-oxide) is aneffective antibacterial agent against a wide variety of microorganisms(bacteria, yeasts, and fungi) and is substantive to skin, paper, andcloth.

EXAMPLE III Shampoo composition Composition sulfated coconut fattyalcohol-sodium Salt 23 23 Sodium stearate 8. 7 8. 7 Sodium alkylglyceryl ether sulfonate 1 28. 3 Sodium acyl sareosinate 2 5. 0 Sodiumsulfate 0.8 0.8 2.6 Sodium chloride 6. 6 Trisodium phosphate 2. 1 2. 1Diethanolamide of coconut fatty acids 2. 0 Acetylated lanolin 1. 0 1.0 1. 0 Perfume 1. 0 1. 0 O. 4 Phenylbismuth bis (2-pyridinethiol 1- 0.5 1. 0 1. 5 Balance Balance Balance 7. 7 7. 7 7. 4

1 Alkyl radicals derived from fatty alcohol, 25.3% from coconut and 3%form tallow 2 Acyl radicals derived from coconut fatty acids.

Composition #4: Percent Dimethyl coconut 1 amine oxide 8.0 Disodiumlauryl beta-iminodipropionate 5.0 Sodium coconut 1 sulfate 4.0 Coconut 2diethanolamide 1.0 Phenylbismuth bis(2-pyridinethiol l-oxide) 1.0Perfume 0.5 Color 0.01

Citric acid to adjust pH to 7.5 Water, balance.

The alkyl radical is derived from middle cut coconut alcohol and hasapproximately the following chain length compo sivtplllg C10, 66% C12,23% C14, and 9% Cm.

Composition #5: A shampoo composition is obtained by uniformly mixingtogether the following ingredients:

3% monoethanolamide of coconut oil fatty acids having the followingchain length distribution: 16% C 48%): C12, 17% C14, 9% C16, and 10% C181% Phenylbismuth bis(2-pyridinethiol l-oxide) 3% diethanolamide ofcoconut oil fatty acids having the following chain length distribution:16% C 48% C12, 117% C14, C16 and C13 7% ethanol 0.75% methyl cellulose,a 2% solution of which has a viscosity of 4000 cps. at 68 F. and a gelpoint of 140 F.

0.75% perfume balance, water.

Shampoo formulations containing phenylbismuth bis(2- pyridinethioll-oxide) are desirable since they will control dandruff.

EXAMPLE IV Personal use detergent lotion Composition #6: PercentPotassium coconut glyceryl ether sulfonate (about 23% diglyceryl and thebalance substantially all monoglyceryl) Sodium coconut glyceryl ethersulfonate (diglyceryl and monoglyceryl content as above) Sodium tallowglyceryl ether sulfonate (diglyceryl and monoglyceryl content as above);the tallow alkyl radicals correspond to those of substantially saturatedtallow alcohols containing approximately 2% C 32% C16, and C13 Coconutdimethylamine oxide Sodium salt of sulfated condensation product of onemole of nonylphenol with 4 moles ethylene oxide Potassium pyrophosphatePhenylbismuth bis(2-pyridinethiol l-oxide) Sodium chloride (in additionto that from detergents) Sodium toluene sulfonate Sodiumcarboxymethylcellulose (degree of substitution 0.65-0.95; viscosity of1% soln., 1000-2800 cps. at 25 C.) Acrylamide polymer (contains 5-10mole percent acrylic acid radicals; monomer is less than 0.05%;viscosity of a 0.5% soln. is about -15 centipoises at 25 C.) Salts(sodium and potassium chloride and sulfate from detergents) 1-2 WaterBalance Coconut: indicates alkyl radicals corresponding to those ofmiddle cut coconut fatty alcohol containing approximately 2% C10, 66%C12, 23% C11, and 9% C15.

EXAMPLE V Cream shampoo Composition: Percent Sodium coconut glyccrylether sulfonate (about 29% diglyceryl and the balance substantiallymonoglyceryl) 14.8 Sodium tallow glyceryl ether sulfonate (about 28%diglyceryl and the balance substantially monoglyceryl) 2.0 Sodiumchloride 6.7 Sodium sulfate 3.5 Sodium N-lauroyl sarcosinate 3.0Phenylbismuth bis(2-pyridinethiol l-oxide) 2.0 Middle-cut coconut 1diethanolamine 0.5 Acetylated lanolin 1.0 Perfume 0.4 Water Balance 1 2%C10, 66% C12, 23% C14, and 9% C10.

12 EXAMPLE VI acids 17.0 Inorganic salts (sodium and potassium chloridesand sulfates) 32.0 Phenylbismuth bis(2-pyridinethiol l-oxide) 1.0 Waterand minors Middle-cut coconut alcohols having a chain length distribution substantially as follows: 2% C10, 66% C12, 23% C14 and 9% C10.

2 Balance to 100.

This bar cleans well and exhibits good odor reducing propertiesevidencing antibacterial eifectiveness. It reduces the number ofbacteria on the skin significantly.

EXAMPLE VII A granular built synthetic detergent composition having thefollowing formulation can be prepared and the antibacterial compositionsof the present invention can be incorporated therein.

Percent Sodium dodecylbenzene sulfonate 17.5 Sodium tripolyphosphate50.0 Sodium sulphate 14.0 Sodium silicate (SiO :Na O=2:1) 7.0Phenylbismuth bis(2-pyridinethiol l-oxide) 3.0 Water and minors 1Balance to 100.

This composition, in addition to performing well in its cleaningcapacity, imparts considerable antibatcerial activity to fabricscleansed in its solution.

Substantially equivalent results are obtained, i.e., good cleaning andgood odor reducing properties when the sodium dodecylbenzene sulfonateof Example VII is replaced, on an equal weight basis, by the following:

dodecyltrimethylammonium chloride; nonylbenzylethyldimethylammoniumnitrate; tetradecylpyridinium bromide;octadecylbutylpropylmethylphosphonium nitrite; decyldimethylsulfom'umchloride; (hexylphenyl)dimethylbenzylammonium fluoride;eicosyldimethylbenzylphosphonium chloride;coconutalkylmethylmorpholininm nitrate; octadecylmethylbenzylsulfoniumsulfate; laurylpyridinium chloride;

lanrylpyridinium bromide;

laurylpyridinium bisulfate;laurylpyridinium-S-chloro-2-mercaptobenzothiazole;laurylpicolinium-p-toluenesulfonate; tetradecylpyridinium bromide;

cetylpyridinium chloride;

cetylpyridinium bromide;

laurylisoquinolinium bromide; laurylisoquinolinium saccharinate;alkylisoquinolinium bromide;

N-cetyl- -ethyl-morpholinium ethosulfate; benzalkonium chloride;

monoquaternaries R N+X* one R group is fatty);octadecyltrimethylammoninm chloride;

coconut alkyl trimethylammonium chloride; dodecylbenzyltn'(octyldecyl)ammonium chloride; monoquaternaries R N+X- (two R groups are fatty);dlhexadecyldimethylammonium chloride; di-coconut alkyl dimethylammoniumchloride;

13 monoquaternaries R.,N+X (three R groups are fatty); tri(hydrogenatedtallow) methylammoniurn chloride; distilled tallow amine acetate;diamine acetates; N-oleyl propylene diamine monoacetate; sodium tallowalkyl sulfate; potassium coconut alkyl glyceryl ether sulfonate; sodiumsalt of randomly sulfonated paraffin containing an average of 15.2carbon atoms; ammonium tridecyl sulfate; condensation product of octylphenol with 15 moles of ethylene oxide per mole of octyl phenol;

dimethyldodecylarnine oxide;

dodecyldimethylphosphine oxide;

tetradecyl methyl sulfoxide;

3- (N,N-dimethyl-N-hexadecylammonio) prop anel-sulfonate;

3-dodecylaminopropionate; and

dodecyl-fl-alanine.

The invention has been described above in conjunction with toilet andlaundry detergents. It will be obvious to those skilled in the art,however, that the antibacterial compositions of the present inventioncan also be beneficially employed in such products as shampoos, footpowders, antiseptic ointments, cosmetic products and the like.

A fabric softening composition having the following formulation can beprepared. The antibacterial and antifungal effectiveness of thephenylbismuth bis(2-pyridinethiol l-oxide) is especially desirable sincethe antibacterial and antifungal compound is extremely substantive tocloth.

Percent Dialkyl dimethylammonium chloride 75% active ingredientdispersed in isopropanol and water, the dialkyl groups beingapproximately 24% hexadecyl, 75% octadecyl and 1% octadecenyl 7 Thecondensation product of 30 moles of ethylene oxide with one mole ofcoconut alcohol 3 Color and perfume 0.3 Phenylbismuthbis(2-pyridinethiol l-oxide) 1.0 Water Balance Twenty p.p.m. of PBDPwere added to a 0.1% aqueous concentration of Downy, a commercialcationic softener containing a ditallowalkyl dimethylammonium chlorideactive. This composition, when used at 27 C. for ten minutes to treatDacron-prebrightened, nylon, muslin, nylon-prebrightened,muslin-prebrightened, and Dacronzcotton (65 :35)-prebrightened clothwhich had been washed with Ivory Snow, a commercial soap powder, andmachine dried, completely controlled the growth of S. aureus, S.faecalis, E. coli, C. albicans, and P. mirabilz's. With similarcompositions containing 5 p.p.m. PBDP, one rinse gave completeprotection against S. aureus and S. faecalis, after three rinses E. coliand C. albicans are stopped, and after four rinses, P. mirabilis isinhibited. With similar compositions containing 30-40 p.p.m. PBDP, onerinse gave protection against P. aeruginosa on muslin, nylon, and theDacronzcotton blend.

What is claimed is:

1. Phenylbismuth bis(2-pyridinethiol l-oxide).

References Cited UNITED STATES PATENTS 3,239,411 3/1966 Leebrick 260-270R 3,247,050 4/1966 Leebrick 260-270 R 3,321,480 5/1967 Schroder 260-270R 3,583,999 6/1971 Damico 260-270 R 3,236,733 2/1966 Karsten 424-2453,705,943 12/1972 Kaufman 260-270 R OTHER REFERENCES Tenenbaum, Am.Perfumer & Cosmetics, vol. 86, 33-7 (1971).

Gosse et al., Jour. Soc. Cosmetic Chemists, vol. 20, 603-13 (1969).

Van der Wyk et al., Jour. Soc. Cosmetic Chemists, vol. 15, pp. 761-8(1964).

DONALD G. DAUS, Primary Examiner US. Cl. X.R.

. UNETED STATES PATENT OFFICE fiERTiFICATE 6F CORREC'HQN Patent No,3r753r99O D ti d August 21., 1 973 Inventor) John Downing Curry It iscertified that error appears in the above-identified patent and thatsaid Letters Patent are hereby corrected as shown below:

E2ol. 1, line 20 after "This" should read -is. w

Col. 1, line 23, after "Same" should read Col. 1, line 48, "Known"should read -known--. Col. 2 lines 9 & l0 "spider-miais" should read-epidermidis Col. 2, line 20, "dematophytes" should read -dermatophytes.Col. 2, line 40, after "leprosy" should read Col. 4, line 64, after"amine" the should be deleted. Col. 6, line 15, "substitutents" shouldread -substituents--.

Col. 'Z, line 50, "phenylbismuth bis(2 pyridinethio l-o xide)"' shouldread phenylbismuth bis(2pyridinethiol l-oxide)-.

Co-l. l0, line 11, "bacterial" should read bacteria-. Col. 10, line L B,"form should read from. Col. 10, line 62, "25% C should read --2% C Col.12, line 34, "sulph e" should read --sul%%te--.

Signed and sealed this 19th day of March 197b,.

( SEAL Attest:

' EDWARD M. FLETCHER, JR. C. MARSHALL DANN Attesting OfficerCommissioner of Patents

